橡胶树未授粉胚珠愈伤组织乳管细胞的鉴定及分化研究  

Identification of Laticifer Cells of Calli Derived from Non-pollinated Ovules of Hevea brasiliensis and Their Differentiation

作　　者：谭德冠[1] 彭晶[1] 付莉莉[1] 孙雪飘[1] 韩冰莹[1] 张家明[1] TAN Deguan;PENG Jing;FU Lili;SUN Xuepiao;HAN Bingying;ZHANG Jiaming(Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural Sciences(CATAS)/Hainan Academy of Tropical Agricultural Resources,CATAS/Key Laboratory of Tropical Crop Biology and Genetic Resources of the Ministry of Agriculture and Rural Affairs,Haikou,Hainan 571101,China)

机构地区：[1]中国热带农业科学院热带生物技术研究所/中国热带农业科学院海南热带农业资源研究院/农业农村部热带作物生物学与遗传资源利用重点实验室,海南海口571101

出　　处：《热带农业科学》2023年第8期50-55,共6页Chinese Journal of Tropical Agriculture

基　　金：海南省自然科学基金项目(No.2019RC296,No.322RC761);海南省重大科技计划项目(No.ZDKJ2021012)。

摘　　要：乳管是橡胶树合成和贮存胶乳的场所。目前尚不清楚橡胶树未授粉胚珠来源的愈伤组织是否存在乳管细胞及其分化情况。本研究通过组织化学法、免疫组织化学法、分子生物学法证实未授粉胚珠愈伤组织中存在乳管细胞。RT-PCR结果显示,小橡胶粒子蛋白(SRPP)、橡胶延伸因子(REF)基因均能从未授粉胚珠愈伤组织cDNA中扩增出,Blast比对发现,它们的序列与已知的橡胶树树皮胶乳表达的基因序列相一致,进一步证实未授粉胚珠愈伤组织存在乳管细胞,且其功能与树皮乳管细胞相似。培养基中添加低浓度的茉莉酸(JA)能促进未授粉胚珠愈伤组织乳管细胞分化,而高浓度的JA会抑制其分化,JA浓度以1 mg/L效果最佳。橡胶树未授粉胚珠愈伤组织乳管细胞分化依赖于基因型。本研究结果为橡胶树未授粉胚珠愈伤组织可作为研究乳管分化机制的模型提供实验依据,对促进乳管分化机制研究具有一定意义。Laticifers are the place for Hevea brasiliensis to synthesize and store latex.It is unclear whether laticifer cells present in the callus derived from non-pollinated ovules of Hevea brasiliensis and the differentiation of laticifer cells.In the present paper,laticifer cells were confirmed to be present in the calli of non-pollinated ovules by histochemical,immunohis-tochemical,and molecular biological methods.RT-PCR results showed that SRPP and REF genes were amplified from the callus cDNA of non-pollinated ovules.Blast results showed that their sequences were consistent with the known ones ex-pressed in rubber tree latex,further confirming that laticifer cells existed in non-pollinated ovule calli,and their functions were similar to those of laticifer cells in the bark.Adding a low concentration of JA in media promoted the differentiation of non-pollinated ovule callus laticifer cells while adding a high concentration in media inhibited their differentiation.The concentration of JA at 1mg/L was optimal.The differentiation of laticifer cells of Hevea brasiliensis non-pollinated ovule calli was genotype-dependent.The results of this study provide an experimental basis for H.brasiliensis unpollinated ovule calllus tissues that can be used as a model to study the mechanism of laticifer differentiation,which is of significance in promoting the study of the one of laticifer differentiation.

关 键 词：橡胶树 未授粉胚珠愈伤组织 乳管细胞分化 茉莉酸 研究模型 

分 类 号：S794.1[农业科学—林木遗传育种]